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Characterisation of glutathione s-transferase activity in Turkish red pine (Pinus brutia ten) : variation in environmentally cold stressed seedlings

Boyoğlu, Seyhan
Plants can not escape from biotic and abiotic stress factors such as, extreme temperatures, high light intensity, drought, UV radiation, heavy metals, and pathogen attack. Plants have versatile defens systems against such stress conditions. In this study, the role of glutathione S-transferases (GSTs) in cold stress conditions were examined. Glutathione S-transferases are the enzymes that detoxify natural and exogenous toxic compounds by conjugation with glutathione. Glutathione, an endogenous tripeptide, is important as reducing agent, nucleophilic scavenger, and alleviate the chemical toxicity in the plants by the reaction of GSTs. Glutathione conjugates can be transported to the vacuoles or apoplast and are generally much less toxic than the parent compounds. In plants there are four distinct families of the soluble GSTs, namely Phi (F), Type I; Zeta (Z), Type II; Tau (U), Type III; Theta (T), Type IV. By contrast with the mammalian families of GST, relatively little is known about the plant GST families. Up to date, there is not any study on GST isolation and characterization from Turkish red pine, in this respect, this study well play a frontier role the future research dealing with this topic. In this study, some properties of Turkish red pine GST activity towards CDNB (1-chloro-2,4 dinitrobenzene) were examined. The average specific activity of Turkish red pine GST towards CDNB was found as 200l50 (MeanlSE, n= 18) nmole/min/mg cytosolic protein. GSTs in cytosol prepared from Turkish red pine needles retained its activity without loss for four weeks at -80?C. The rate of conjugation reactions were linear up to 0.8mg of Turkish red pine cytosolic protein and 0.4 mg cytosolic protein was routinely used. The Turkish red pine GST showed its maximum activity at pH 8.0 in 25 mM phosphate buffer and 42 C. The measurements were carried out at room temperature (RT) of 25